Biomedical subjects

Martin Hansen

Publications and source records attributed to Martin Hansen.

13 recordsLinked to original sources

PubMed2006

Evidence for a pre-latent form of the serpin plasminogen activator inhibitor-1 with a detached beta-strand 1C.

Latency transition of plasminogen activator inhibitor-1 (PAI-1) occurs spontaneously in the absence of proteases and results in stabilization of the molecule through insertion of its reactive center loop (RCL) as a strand in beta-sheet A and detachment of beta-strand 1C (s1C) at the C-terminal hinge of the RCL. This is one of the largest structural rearrangements known for a folded protein domain without a concomitant change in covalent structure. Yet, the sequence of conformational changes during latency transition remains largely unknown. We have now mapped the epitope for the monoclonal antibody H4B3 to the cleft revealed upon s1C detachment and shown that H4B3 inactivates recombinant PAI-1 in a time-dependent manner. With fluorescence spectroscopy, we show that insertion of the RCL is accelerated in the presence of H4B3, demonstrating that the loss of activity is the result of latency transition. Considering that the epitope for H4B3 appears to be occluded by s1C in active PAI-1, this finding suggests the existence of a pre-latent conformation on the path from active to latent PAI-1 characterized by at least partial detachment of s1C. Functional characterization of mutated PAI-1 variants suggests that a salt-bridge between Arg273 and Asp224 may stabilize the pre-latent conformation. The binding of H4B3 and of a peptide targeting the cleft revealed upon s1C detachment was hindered by the glycans attached to Asn267. Conclusively, we have provided evidence for the existence of an equilibrium between active PAI-1 and a pre-latent form, characterized by reversible detachment of s1C and formation of a glycan-shielded cleft in the molecule.

Animals↗

PubMed2006

Objectively measured and subjectively perceived distortion in nonlinear systems.

A method for measuring nonlinear distortion, which is applicable to almost any transmission system and which can use any input signal as a test signal, is proposed. The method exploits the fact that a pair of signals, generated to form the real and imaginary parts of the analytical signal corresponding to the input signal to a system, lose their property of being a Hilbert pair after being passed through a nonlinear system. The method was tested by measuring 12 different hearing aid compression systems. These objective measurements were compared with the subjectively perceived amount of distortion, assessed by a group of 10 otologically normal subjects. A reasonably monotonic relation between the subjective and objective measures of distortion was observed. The objective Hilbert pair based measure can be related to both traditional total harmonic distortion and traditional signal to noise ratio.

Journal Article↗

PubMed2005

A urokinase-type plasminogen activator-inhibiting cyclic peptide with an unusual P2 residue and an extended protease binding surface demonstrates new modalities for enzyme inhibition.

To find new principles for inhibiting serine proteases, we screened phage-displayed random peptide repertoires with urokinase-type plasminogen activator (uPA) as the target. The most frequent of the isolated phage clones contained the disulfide bridge-constrained sequence CSWRGLENHRMC, which we designated upain-1. When expressed recombinantly with a protein fusion partner, upain-1 inhibited the enzymatic activity of uPA competitively with a temperature and pH-dependent K(i), which at 25 degrees C and pH 7.4 was approximately 500 nm. At the same conditions, the equilibrium dissociation constant K(D), monitored by displacement of p-aminobenzamidine from the specificity pocket of uPA, was approximately 400 nm. By an inhibitory screen against other serine proteases, including trypsin, upain-1 was found to be highly selective for uPA. The cyclical structure of upain-1 was indispensable for uPA binding. Alanine-scanning mutagenesis identified Arg(4) of upain-1 as the P(1) residue and indicated an extended binding interaction including the specificity pocket and the 37-, 60-, and 97-loops of uPA and the P(1), P(2), P(3)', P(4)', and the P(5)' residues of upain-1. Substitution with alanine of the P(2) residue, Trp(3), converted upain-1 into a distinct, although poor, uPA substrate. Upain-1 represents a new type of uPA inhibitor that achieves selectivity by targeting uPA-specific surface loops. Most likely, the inhibitory activity depends on its cyclical structure and the unusual P(2) residue preventing the scissile bond from assuming a tetrahedral geometry and thus from undergoing hydrolysis. Peptide-derived inhibitors such as upain-1 may provide novel mechanistic information about enzyme-inhibitor interactions and alternative methodologies for designing effective protease inhibitors.

4-Aminobenzoic Acid↗

PubMed2005

Conversion of proteins from a non-polarized to an apical secretory pattern in MDCK cells.

Previously it was shown that fusion proteins containing the amino terminus of an apical targeted member of the serpin family fused to the corresponding carboxyl terminus of the non-polarized secreted serpin, antithrombin, are secreted mainly to the apical side of MDCK cells. The present study shows that this is neither due to the transfer of an apical sorting signal from the apically expressed proteins, since a sequence of random amino acids acts the same, nor is it due to the deletion of a conserved signal for correct targeting from the non-polarized secreted protein. Our results suggest that the polarity of secretion is determined by conformational sensitive sorting signals.

Amino Acid Sequence↗

PubMed2005

Assessment of the importance of sorption for steroid estrogens removal during activated sludge treatment.

Distribution coefficients (K(d)) between water and activated sludge particles (f(oc)=27.7+/-0.1%) were measured for the steroid estrogens (SE), estrone (E1), 17beta-estradiol (E2) and 17alpha-ethinylestradiol (EE2) in batch experiments. Experimental concentration levels ranged from environmentally realistic low ng/l to the high microg/l. In this range K(d)s were independent of their water concentration. The experimentally obtained K(d)s (with 95% confidence intervals) were 402+/-126 l/kg, 476+/-192 l/kg and 584+/-136 l/kg for E1, E2 and EE2, respectively. K(d)s were used to estimate the fraction of the total SE concentration that is expected to be sorbed in the activated sludge treatment tanks of a typical STP assuming equilibrium conditions. Assuming a suspended solids concentration of 4 g/l dissolved solids (ds), it was estimated that 61+/-9%, 66+/-13% and 70+/-6% of the total concentration of E1, E2 and EE2, respectively, would be sorbed during activated sludge treatment. The fraction of the SEs that was expected to be sorbed to suspended sludge particles in the effluents from a typical Danish STP was estimated to be only 0.20+/-0.06%, 0.24+/-0.10% and 0.29+/-0.07% of the total concentration of E1, E2 and EE2, respectively, at a suspended solids concentration of 5 mg/lds. For a typical STP the removal of steroid estrogens with excess sludge was estimated to be only 1.5-1.8% of the total loading if equilibrium conditions exists. Sorption is therefore not important for the fate of SEs in STPs compared to biodegradation.

Biodegradation, Environmental↗

PubMed2005

Ultrasound research scanner for real-time synthetic aperture data acquisition.

Conventional ultrasound systems acquire ultrasound data sequentially one image line at a time. The architecture of these systems is therefore also sequential in nature and processes most of the data in a sequential pipeline. This often makes it difficult to implement radically different imaging strategies on the platforms and makes the scanners less accessible for research purposes. A system designed for imaging research flexibility is the prime concern. The possibility of sending out arbitrary signals and the storage of data from all transducer elements for 5 to 10 seconds allows clinical evaluation of synthetic aperture and 3D imaging. This paper describes a real-time system specifically designed for research purposes. The system can acquire multichannel data in real-time from multi-element ultrasound transducers, and can perform some real-time processing on the acquired data. The system is capable of performing real-time beamforming for conventional imaging methods using linear, phased, and convex arrays. Image acquisition modes can be intermixed, and this makes it possible to perform initial trials in a clinical environment with new imaging modalities for synthetic aperture imaging, 2D and 3D B-mode, and velocity imaging using advanced coded emissions. The system can be used with 128-element transducers and can excite 128 transducer elements and receive and sample data from 64 channels simultaneously at 40 MHz with 12-bit precision. Two-to-one multiplexing in receive can be used to cover 128 receive channels. Data can be beamformed in real time using the system's 80 signal processing units, or it can be stored directly in RAM. The system has 16 Gbytes RAM and can, thus, store more than 3.4 seconds of multichannel data. It is fully software programmable and its signal processing units can also be reconfigured under software control. The control of the system is done over a 100-Mbits/s Ethernet using C and Matlab. Programs for doing, e.g., B-mode imaging can be written directly in Matlab and executed on the system over the net from any workstation running Matlab. The overall system concept is presented along with its implementation and examples of B-mode and in vivo synthetic aperture flow imaging.

Analog-Digital Conversion↗

PubMed2004

Plasminogen activator inhibitor-I and tumour growth, invasion, and metastasis.

In recent decades, evidence has been accumulating showing the important role of urokinase-type plasminogen activator (uPA) in growth, invasion, and metastasis of malignant tumours. The evidence comes from results with animal tumour models and from the observation that a high level of uPA in human tumours is associated with a poor patient prognosis. It therefore initially came as a surprise that a high tumour level of the uPA inhibitor plasminogen activator inhibitor-I (PAI-I) is also associated with a poor prognosis, the PAI-I level in fact being one of the most informative biochemical prognostic markers. We review here recent investigations into the possible tumour biological role of PAI-I, performed by animal tumour models, histological examination of human tumours, and new knowledge about the molecular interactions of PAI-I possibly underlying its tumour biological functions. The exact tumour biological functions of PAI-I remain uncertain but PAI-I seems to be multifunctional as PAI-I is expressed by multiple cell types and has multiple molecular interactions. The potential utilisation of PAI-I as a target for anti-cancer therapy depends on further mapping of these functions.

Animals↗

PubMed2003

Mapping of the epitope of a monoclonal antibody protecting plasminogen activator inhibitor-1 against inactivating agents.

Plasminogen activator inhibitor-1 (PAI-1) belongs to the serpin family of serine proteinase inhibitors. Serpins inhibit their target proteinases by an ester bond being formed between the active site serine of the proteinase and the P1 residue of the reactive centre loop (RCL) of the serpin, followed by insertion of the RCL into beta-sheet A of the serpin. Concomitantly, there are conformational changes in the flexible joint region lateral to beta-sheet A. We have now, by site-directed mutagenesis, mapped the epitope for a monoclonal antibody, which protects the inhibitory activity of PAI-1 against inactivation by a variety of agents acting on beta-sheet A and the flexible joint region. Curiously, the epitope is localized in alpha-helix C and the loop connecting alpha-helix I and beta-strand 5A, on the side of PAI-1 opposite to beta-sheet A and distantly from the flexible joint region. By a combination of site-directed mutagenesis and antibody protection against an inactivating organochemical ligand, we were able to identify a residue involved in conferring the antibody-induced conformational change from the epitope to the rest of the molecule. We have thus provided evidence for communication between secondary structural elements not previously known to interact in serpins.

Antibodies, Monoclonal↗

PubMed2003

Ensuring accuracy of the pediatric hearing aid fitting.

Considerable effort has recently been dedicated towards early detection of hearing loss in infants. The goal is to subsequently fit hearing aids to hearing-impaired infants so that they can develop speech and language as early and as well as possible. The responsibility of the audiologist is to provide the child with the optimal amplification from the hearing aid. This paper gives an overview of aspects involved in correctly assessing the hearing loss and optimally fitting and verifying the performance of the child's hearing aids. Special emphasis is directed towards differences that exist between children and adults, and how these differences can be considered during a pediatric fitting. We suggest a stringent terminology that can help avoid ambiguous terms and connotations in the child's domain which are derived from those that have become established in adult audiometry and hearing aid fittings. The quantification of hearing thresholds and their appropriate representation is reviewed. An appropriate threshold-based prescription of gain or output of the hearing aid is described. Issues of verification and the actual programming of the hearing aid are discussed.

Journal Article↗

PubMed2002

RunMACRO: a program to facilitate sensitivity analyses using the water flow and solute movement model MACRO.

Sensitivity analyses of pesticide leaching often involve a large range of simulations based on nearly identical set-ups. Using RunMACRO it is possible to make large numbers of simulations with a minimum of exertion. Running many nearly identical model set-ups is tedious and might introduce errors in selecting the correct values from a long range of parameter files that are appropriate to the model set-up. RunMACRO makes the task easier and minimises the risk of errors in the generation of parameter files and model executions. Using RunMACRO, it is possible to create a suite of MACRO simulations based on a single parameter file where the range or a list of values for one to three parameters is specified. RunMACRO then creates a parameter file for each of the simulations and runs the simulations one by one. RunMACRO can easily be modified to be used with other simulation programs that use ASCII-based parameter files and can be started from a command prompt. RunMACRO is available free to use and modify from the Geological Survey of Denmark and Greenland's home page.

Algorithms↗

PubMed2002

Effects of multi-channel compression time constants on subjectively perceived sound quality and speech intelligibility.

OBJECTIVE: The purpose of this study was to determine the influence of the compression time constants in a multi-channel compression hearing aid on both subjectively assessed speech intelligibility and sound quality in realistic binaural acoustical situations for normal-hearing and hearing-impaired listeners. DESIGN: A nonlinear hearing aid with 15 independent compression channels of approximated critical bandwidth was simulated on a personal computer. Various everyday life situations containing different sounds such as speech and speech in noise were recorded binaurally through original hearing aid microphones placed in BTE hearing aid cases. Two experiments were run with normal hearing and hearing-impaired subjects. For each subject, hearing thresholds were established using in situ audiometry. The static I/O-curve parameters in all channels of the hearing aid were then adjusted so that normal speech received an insertion gain corresponding to the NAL-R formula (Byrne & Dillon, 1986). The compression ratio was kept constant at 2.1:1. In the first experiment with six normal-hearing and six hearing-impaired subjects, the hearing aid was programmed to four different settings by changing only the compression time constants while all the parameters describing the static nonlinear Input/Output-curve were kept constant. The compression threshold was set to a very low value. In the second experiment with seven normal-hearing and eight hearing-impaired subjects, the hearing aid was programmed to four settings by changing the release time constants and the compression threshold while all other remaining parameters were kept constant. Using a complete A/B pair comparison procedure, subjects were presented binaurally with the amplified sounds and asked to subjectively assess their preference for each hearing aid setting with regards to speech intelligibility and sound quality. RESULTS AND CONCLUSIONS: In Experiment 1, all subjects showed a significant preference for the longest release time (4 sec) over the two shorter release times (400 msec and 40 msec), pertaining to quality and intelligibility. In combination with the long release time, the attack time (1 msec or 100 msec) was of less importance. Larger inter-individual differences were observed among all subjects with regard to the perceived sound quality of musical and nonspeech signals. In Experiment 2 the hearing-impaired subjects showed a significant preference for the hearing aid setting with a long release time (4 sec) and a low compression threshold (20 dB SPL), both with regard to sound quality and speech intelligibility. With a short release time (40 msec), a lower compression threshold (20 dB SPL) was preferred over a higher threshold (50 dB SPL). Among normal-hearing subjects, the same settings were also preferred for speech intelligibility. However, no significant differences were observed in these subjects regarding the assessment of sound quality. Again, more individual variability was found during the assessment of music and nonspeech signals. In sum, the preference of a rather long release time raises the question of whether a compression system with a very short time constant is the optimal means to compensate for the sensory recruitment phenomenon.

Acoustic Stimulation↗

PubMed2002

The molecular basis for anti-proteolytic and non-proteolytic functions of plasminogen activator inhibitor type-1: roles of the reactive centre loop, the shutter region, the flexible joint region and the small serpin fragment.

The serine proteinase inhibitor plasminogen activator inhibitor type-1 (PAI-1) is the primary physiological inhibitor of the tissue-type and the urokinase-type plasminogen activator (tPA and uPA, respectively) and as such an important regulator of proteolytic events taking place in the circulation and in the extracellular matrix. Moreover, a few non-proteolytic functions have been ascribed to PAI-1, mediated by its interaction with vitronectin or the interaction between the uPA-PAI-1 complex bound to the uPA receptor and members of the low density lipoprotein receptor family. PAI-1 belongs to the serpin family, characterised by an unusual conformational flexibility, which governs its molecular interactions. In this review we describe the anti-proteolytic and non-proteolytic functions of PAI-1 from both a biological and a biochemical point of view. We will relate the various biological roles of PAI-1 to its biochemistry in general and to the different conformations of PAI-1 in particular. We put emphasis on the intramolecular rearrangements of PAI-1 that are required for its antiproteolytic as well as its non-proteolytic functions.

Animals↗

PubMed2001

A single, bi-functional aquaglyceroporin in blood-stage Plasmodium falciparum malaria parasites.

The malaria parasite Plasmodium falciparum faces drastic osmotic changes during kidney passages and is engaged in the massive biosynthesis of glycerolipids during its development in the blood-stage. We identified a single aquaglyceroporin (PfAQP) in the nearly finished genome of P. falciparum with highest similarity to the Escherichia coli glycerol facilitator (50.4%), but both canonical Asn-Pro-Ala (NPA) motifs in the pore region are changed to Asn-Leu-Ala (NLA) and Asn-Pro-Ser (NPS), respectively. Expression in Xenopus oocytes renders them highly permeable for both water and glycerol. Sugar alcohols up to five carbons and urea pass the pore. Mutation analyses of the NLA/NPS motifs showed their structural importance, but the symmetrical pore properties were maintained. PfAQP is expressed in blood-stage parasites throughout the development from rings via trophozoites to schizonts and is localized to the parasite but not to the erythrocyte cytoplasm or membrane. Its unique bi-functionality indicates functions in the protection from osmotic stress and efficiently provides access to the serum glycerol pool for the use in ATP generation and primarily in the phospholipid synthesis.

Adenosine Triphosphate↗